Hematologic lineages in MPLW515L/JAK2V617F double-mutated Ph− CMPD
| Type of blood or bone marrow cell . | Source . | Mode of separation . | MPLW515L (% mutant alleles)‡ . | JAK2V617F (% mutant alleles)‡ . |
|---|---|---|---|---|
| Mononuclear cells | PB | DGC | 52 | 14 |
| B lymphocytes (CD19+, purity ∼95%) | PB | FACS | 37 | <1 |
| T lymphocytes (CD3+, purity ∼95%) | PB | FACS | 52 | <1 |
| CD34+ cells (∼300 cells) | PB | FACS | 94 | 44 |
| Granulocytes (CD14−/CD15+/CD16+, purity ∼95%) | PB | FACS | 47 | 62 |
| Monocytes (CD14+, purity ∼95%) | PB | FACS | 69 | 53 |
| Eosinophils* (∼100 cells) | PB | LMD | <1 | 92 |
| Total bone marrow cells | BM | 58 | 57 | |
| Erythropoieis† (hemoglobin-positive, ∼100 cells) | BM | LMD | <1 | 58 |
| Megakaryocytes† (∼100 cells) | BM | LMD | <1 | 30 |
| Type of blood or bone marrow cell . | Source . | Mode of separation . | MPLW515L (% mutant alleles)‡ . | JAK2V617F (% mutant alleles)‡ . |
|---|---|---|---|---|
| Mononuclear cells | PB | DGC | 52 | 14 |
| B lymphocytes (CD19+, purity ∼95%) | PB | FACS | 37 | <1 |
| T lymphocytes (CD3+, purity ∼95%) | PB | FACS | 52 | <1 |
| CD34+ cells (∼300 cells) | PB | FACS | 94 | 44 |
| Granulocytes (CD14−/CD15+/CD16+, purity ∼95%) | PB | FACS | 47 | 62 |
| Monocytes (CD14+, purity ∼95%) | PB | FACS | 69 | 53 |
| Eosinophils* (∼100 cells) | PB | LMD | <1 | 92 |
| Total bone marrow cells | BM | 58 | 57 | |
| Erythropoieis† (hemoglobin-positive, ∼100 cells) | BM | LMD | <1 | 58 |
| Megakaryocytes† (∼100 cells) | BM | LMD | <1 | 30 |
The 67-year-old female patient presented with splenomegaly and leukocytosis (hemoglobin 151 g/L; leukocytes 13.6 × 109/L; eosinophils 0.4-0.65 × 109/L; platelets 136 × 109/L; no blasts). Cytogenetic analysis revealed trisomy 9 [47, XX, +9, 6/47, idem, del (13)(q13q21) 5/46,XX 4. nuc ish 9q34 (ABL × 3), 22q11(BCR × 2) 78/100].
BM indicates bone marrow; DGC, density gradient centrifugation; FACS, fluorescence-activated cell sorting (FACS; FACSAria, Becton Dickinson, NJ); LMD, laser microdissection (LMD, SmartCutPlus System based on a CKX41 inverse microscope, Olympus, Hamburg, Germany); and PB, peripheral blood.
Isolated from May-Grünwald-Giemsa–stained cytospins (Figure S2).
Isolated from bone marrow trephine sections.
Allele burden was determined by pyrosequencing (Biotage, Uppsala, Sweden) as recently described.5